ACAT inhibitory agent

ABSTRACT

PCT No. PCT/JP92/00857 Sec. 371 Date Feb. 1, 1993 Sec. 102(e) Date Feb. 1, 1993 PCT Filed Jul. 6, 1992 PCT Pub. No. WO93/02185 PCT Pub. Date Feb. 4, 1993An ACAT inhibitory method and composition containing a monoacylglycerol (monoglyceride) as an active ingredient.

TECHNICAL FIELD

This application is a 371 of PCT/JP92/00857 filed Jul. 06, 1992.

The present invention relates to a novel inhibitory agent for an ACAT enzyme, and more specifically, it relates to a lipid absorption inhibitory edible composition, etc., containing an inhibitory agent for an ACAT enzyme derived from a food component.

BACKGROUND ART

The absorption of a lipid, particularly cholesterol, is believed to be rate determined by a process wherein free cholesterol is converted to ester cholesterol by an ACAT (AcylCoA-Cholesterolacyltransferase) in small intestine epithelial cells. An ACAT inhibitory agent has been found to be useful as an anti-cholesterol agent in this respect. At present, several of these agents are being developed as pharmaceuticals.

DISCLOSURE OF THE INVENTION

As a result of earnest efforts pertaining to the above, the inventors of the present invention found that monoacylglycerol (monoglyceride), a certain kind of fatty acid-like substance derived from microorganism lipids, such as that from lactic acid bacteria and decomposition products of rice bran and wheat bran lipid, demonstrates remarkable inhibitory activity against said enzyme, as those suitable for common use as a so-called functional food component and so forth, and, as a result, the present invention has completed.

Accordingly, in consideration of the present state of the above-mentioned prior art, an object of the present invention is to provide a compound having a remarkable ACAT inhibitory activity, despite being derived from edible natural substances and being essentially non-toxic, as well as an edible composition containing said compound as the active ingredient.

In accordance with the present invention, there are provided an ACAT inhibitory agent containing monoacylglycerol (monoglyceride) as an active ingredient, and a lipid absorption inhibitory edible composition containing the ACAT inhibitory agent and an edible carrier. Although there is no particular limitation to the content of monoacylglycerol (monoglyceride) in the composition, a content of 0.5 to 3% by weight is preferable. Specific examples of the edible carrier include hydroxypropylmethyl cellulose phthalate and casein.

BEST MODE FOR CARRYING OUT THE INVENTION

The present invention will now be explained in detail.

Fatty Acid-like Substance

Those present in microorganism lipids such as those of lactic acid bacteria, and hydrolyzed (enzyme) decomposition products of vegetable lipids such as rice bran and wheat bran lipids, which is purified and isolated in a conventional manner, or commercially available monoacylglycerols (monoglycerides) having 22 carbon atoms or less, preferably 16 carbon atoms or less, such as 2-oleylglycerol, 1-oleylglycerol, 1-linoleylglycerol, 1-linolenoylglycerol, 1-stearoylglycerol, 1-palmitoylglycerol or 1-palmitooleylglycerol, exhibit fatty acid-like properties.

Method of Use and Dosage

The above-mentioned substances can usually be orally administered in a dosage of about 0.1-10 mg/kg (body weight) per day in ordinary form such as tablet or capsule. Since these substances are food components, they are essentially non-toxic when taken orally. On the other hand, in the case of providing same in the form of an edible composition, these substances need not be particularly isolated for utilization, and may be utilized as food auxiliary components such as a microorganism lipid, or the lipid decomposition products of rice bran or wheat bran.

Method for Assaying ACAT Activity

An ACAT activity was assayed according to the method of Heider et al (J. Lipid. Res. 24, 1127-1134 (1983)). That is, a rabbit is fed a 1% cholesterol diet for 3 weeks followed by dissection and extraction of the small intestine. After peeling off the small intestine epithelium and crushing with a homogenizer, incompletely crushed solids are removed by centrifugal separation. The microsome fraction of the 107,000× g sediment was suspended in a 0.154M phosphate buffer (pH 7.4), which is then used for ACAT enzyme preparation. The reaction is carried out by adding 18 nmol of bovine serum albumin and 18 nmol of [1- ¹⁴ C]oleylCoA (0.018 μCi/nmol) to 0.5 ml of a 0.154M phosphate buffer (pH 7.4), heating for 5 minutes at 37° C. and adding 40 μl of the above-mentioned ACAT enzyme preparation (0.2-0.4 mg as protein). After allowing the reaction to proceed for the prescribed amount of time, 7 ml of a mixture of dichloromethane: methanol (2:1) is added to the reaction mixture and thoroughly mixed. The reaction, is stopped and the dichloromethane layer is removed, after which the cholesterol oleate portion is separated from that fraction by silica gel TLC (developing solution: petroleum ether:diethyl ether=95:5). The separated portion is scraped off and the radioactivity is measured using a liquid scintillation counter to calculate ACAT activity.

Preparation of Monoacylglycerol (Monoglyceride)

The monoacylglycerol (monoglyceride), such as 2-oleylglycerol, 1-oleylglycerol, used in the present invention are known substances that are, for example, commercial reagents available from the Sigma Corp. (the compounds of the present invention used in the following Examples are all reagents of the Sigma Corp.).

EXAMPLES

The present invention will now be further illustrated by, but is by no means limited to, the following Examples.

(1) ACAT Inhibitory Activity of Fatty Acid-Like Substance Monoglycerides

(1) In Test Tube

                  TABLE 1                                                          ______________________________________                                                    Inhibitory Ratio (%)                                                           Concentration (μg/ml)                                            Monoglycerides                                                                              200      150    100    50  20                                     ______________________________________                                         2-oleylglycerol                                                                             100      94     82     62  32                                     1-oleylglycerol                                                                             100      92     81     59  30                                     1-linoleylglycerol                                                                          93       86     74     54  28                                     1-linolenoylglycerol                                                                        92       86     76     50  25                                     1-stearoylglycerol                                                                          49       45     38     29  15                                     1-palmitoylglycerol                                                                         94       86     75     56  26                                     1-palmitooleylglycerol                                                                      91       84     72     53  24                                     ______________________________________                                    

(2) Animal Test

A test substance according to the present invention was administered to hamsters (males, body weight: 100 g) at a dose of 1 mg/animal/day by mixing in the same to their feed. The ACAT activity of small intestine epithelial cells was then measured after one week and after two weeks. The results are indicated in Table 2.

                  TABLE 2                                                          ______________________________________                                                       ACAT Inhibitory Ratio (%)                                        Monoglycerides  After 1 week                                                                              After 2 weeks                                       ______________________________________                                         2-oleylglycerol 64         56                                                  1-oleylglycerol 60         52                                                  1-linoleylglycerol                                                                             59         53                                                  1-linolenoylglycerol                                                                           57         51                                                  1-stearoylglycerol                                                                             31         29                                                  1-palmitoylglycerol                                                                            56         50                                                  1-palmitooleylglycerol                                                                         57         54                                                  ______________________________________                                    

INDUSTRIAL APPLICABILITY

As mentioned in detail above, the ACAT inhibitory agent of the present invention containing a monoacylglycerol (monoglyceride) as an active ingredient is substantially non-toxic since it is derived from edible natural substances. Moreover, since it demonstrates remarkable ACAT inhibitory activity, it can be suitably used in functional food components and so forth. 

We claim:
 1. An ACAT inhibitory composition containing as an active ingredient from 0.5 to 3% by weight, based on the total weight of the composition, of a monoacylglycerol selected from the group consisting of 1-stearoylglycerol, 1-palmitoylglycerol and a mixture thereof and an acceptable carrier therefor.
 2. A lipid absorption inhibitory edible composition containing an ACAT inhibitory amount of from 0.5 to 3% by weight, based on the total weight of the composition, of a monoacylglycerol selected from the group consisting of 1-stearoylglycerol, 1-palmitoylglycerol and a mixture thereof and an edible carrier therefor.
 3. A method for inhibiting the absorption of lipids in persons, which comprises administering to a person in need thereof an ACAT inhibitory composition containing as the active ingredient from 0.5 to 3% by weight, based on the total weight of the composition, of a monoacylglycerol selected from the group consisting of 1-stearoylglycerol, 1-palmitoylglycerol and a mixture thereof and an acceptable carrier therefor. 